This project integrates two complementary multiplex, high-throughput analytical protocols on the Luminex xMAP platform - for both immunoassay and highly multiplexed DNA mutation analysis by suspension array - requiring new yet practical investment in technology by newborn screening (NBS) programs. The objective of this proposal is to investigate opportunities to integrate methodologies with this new combined Luminex testing system to develop a comprehensive NBS platform. The project will develop comprehensive assays on the Luminex xMAP platform for use in NBS programs. Two development paths will be described: DNA mutation analysis - using the Luminex liquid universal array platform to develop assays for galactosemia (5 mutations); MCADD (5 mutations), biotinidase deficiency (5 mutations); hearing loss due to connexin-26 in the GJB2 gene (3 mutations) (35); and cytomegalovirus virus (3 markers). Phenotypic analysis - using mUltiplex microsphere sets with conjugated antibodies for specific analyte detection to screen for congenital hypothyroidism, cystic fibrosis, congenital adrenal hyperplasia, and the leukodystrophies Krabbe and Pompe disease.